System and method for automatic plant tissue sampling

ABSTRACT

An automatic plant tissue sampler and a method for operating the same are provided. The sampler can include a plant handler configured to transport a plurality of plants to an imager. The imager may be configured to image plants to identify a sampling location. The automatic plant tissue sampler also includes a sampler configured to remove a tissue sample from the sampling location of plants, and a collection vessel configured to receive the tissue samples. The automatic plant tissue sampler may transport a plurality of plants to an imager and images the plurality of plants to identify a sampling location. The automatic plant tissue sampler can remove a tissue sample from the sampling location of the plurality of plants and store the tissue samples in a collection vessel for testing.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of application U.S. application Ser.No. 14/443,056, filed May 14, 2015, which is a 371 National Stage Entryof PCT/US2013/071929, filed Nov. 26, 2013, which claims benefit of U.S.provisional application No. 61/729,956, filed Nov. 26, 2012,incorporated herein by reference in their entirety.

FIELD OF THE INVENTION

The invention relates generally to plant genetic testing. In particular,the invention relates to a system, method, and apparatus toautomatically sample plant tissue.

DESCRIPTION OF RELATED ART

Development of novel, desirable plant germplasm begins with the analysisand definition of problems and weaknesses of the current germplasm, theestablishment of program goals, and the definition of specific breedingobjectives. The goal is to combine in a single variety beneficiarycombinations of desirable traits from parental germplasm. Development ofsuch combinations may be assisted by testing of the developinggermplasm. Some tests require removal of a tissue sample from thegermplasm to conduct the test. Procurement of tissue samples for thesetests has typically been conducted in a time-consuming manual process.The time-consuming nature of the testing is underscored by the largesample sizes often involved.

SUMMARY

Problems in the prior art are generally solved or circumvented, andtechnical advantages are generally achieved, by embodiments that providea system, method, and apparatus to automatically sample plant tissue asfollows.

According to an illustrative embodiment, an automatic plant tissuesampling system is described. The system includes a plant handlerconfigured to transport one or more plants from a first location to asecond location, and an imager configured to image a plant moved by theplant handler to identify a sampling location. The system also includesa processor in communication with the imager and configured to receivean image of the plant and further configured to select a location on theplant to sample. The system further includes a sampler configured toremove a tissue sample from the sampling location of the plant selectedby the processor, and a collection vessel configured to receive tissuesamples.

In some embodiments, the plant handler is configured to transport theplant from the imager after removal of a tissue sample.

In some embodiments, the plant handler includes a scanner configured toread an identifier associated with the plant, and the collection vesselassociates the tissue sample with the identifier of the sampled plant.

In some embodiments, the scanner is a bar code scanner and theidentifier is a bar code. In another embodiment, the automatic planttissue sampling system further includes a control system configured tostore the location of the tissue sample in the collection vessel in amemory location associated with the bar code of the sampled plant.

In some embodiments, the imager is configured to determine the samplinglocations based on at least one of a size, shape, or color of plantsbeing sampled, or a portion thereof.

In some embodiments, the sample location is a green portion of a leaf.

In some embodiments, the sample location is a rectangular shaped portionof a leaf.

In some embodiments, the sample location is a leaf having a length ofabout 6 mm and width of about 12 mm.

In some embodiments, the imager includes a camera.

In some embodiments, the camera includes an infrared camera, anultraviolet camera, or a visible light camera.

In some embodiments, the sampling location includes a leaf.

In some embodiments, the sampler includes a laser cutter.

In some embodiments, the laser cutter is guided to the sampling locationin response to an image produced by the imager.

In some embodiments, the collection vessel maintains the tissue samplesfor testing.

In some embodiments, the collection vessel includes a microtiterconfigured to receive the tissue sample and associate the tissue samplewith the sampled plant.

In some embodiments, the system includes automatic controlscommunicatively coupled to the plant handler, the imager, and thesampler for operation thereof.

In some embodiments, the processor is in communication with the planthandler and configured to move the plant to the imager.

In some embodiments, the collection vessel is further configured tostore the tissue sample for testing.

According to another illustrative embodiment, a method for samplingplant tissue is described. The method transports one or more plants froma first location to a second location with a plant handler, the secondlocation proximate to an imager. The method images a plant with animager to identify a sampling location, and removes a tissue sample fromthe sampling location of the plant with a sampler. The method stores thetissue sample in a collection vessel. At least the step of imaging aplant and the step of removing a tissue sample are automated.

In some embodiments, the step of imaging a plant includes identifying atleast one of a size, shape, or color of a plurality of plants.

In some embodiments, the method includes identifying the samplinglocation based on at least one of a size, shape, or color of a pluralityof plants.

In some embodiments, the step of imaging the plant includes using avisible light camera, an infrared camera, or an ultraviolet camera togenerate an image.

In some embodiments, the step of transporting one or more plants from afirst location to a second location includes reading an identifierassociated with each plant with a scanner at the first location andtransporting the scanned plant to the second location.

In some embodiments, the step of storing the tissue samples in acollection vessel includes associating the tissue samples with anidentifier of the sampled plant.

In some embodiments, the identifier is a bar code, and the method readsthe bar code with a bar code scanner and stores the location of thetissue sample in the collection vessel in a memory location associatedwith the bar code.

In some embodiments, the method includes transporting the one or moreplants from the imager after removal of a tissue sample.

In some embodiments, the step of removing a tissue sample from thesampling location of the plant includes removing a tissue sample from aleaf of the plant.

In some embodiments, the step of removing a tissue sample from thesampling location of the plant includes removing the tissue sample witha laser cutter.

In some embodiments, the method further includes guiding the lasercutter to the sampling location in response to an image produced by theimager.

In some embodiments, the step of storing the tissue samples in acollection vessel includes maintaining the tissue samples in thecollection vessel for testing.

In still another illustrative embodiment, an automatic plant samplingsystem for automatically sampling plant tissue of an individual plantselected from a group of one or more plants collectively conveyed to theautomatic plant sampling system is described. The automatic plantsampling system includes a frame assembly having a first axis, a secondaxis orthogonal to the first axis, and a third axis orthogonal to thefirst axis and the second axis. The system also includes a planthandling system coupled to the frame assembly so that a portion of theplant handling system moves relative to the frame assembly. The planthandling system is configured to select the individual plant from theone or more plants, transport the individual plant to a sampling systemfor sampling, and transport the individual plant from the samplingsystem to a post sampling location. The sampling system is coupled tothe frame assembly proximate to the plant handling system and configuredto determine a suitable sampling location of the individual plant,sample a portion of plant tissue from the sampling location to create atissue sample, and transport the tissue sample to a storage system. Thestorage system can be coupled to the frame assembly proximate to thesampling system and configured to associate the tissue sample with theindividual plant and maintain the tissue sample in a suitable conditionfor testing. The system further includes a controls systemcommunicatively coupled to the plant handling system, the samplingsystem, and the storage system. The controls system is automaticallyoperable to coordinate movement and operation of the plant handlingsystem, the sampling system, and the storage system for sampling of theindividual plant.

In some embodiments, the plant handling system further includes a plughandler mechanically coupled to the frame assembly. The plug handler isconfigured to move parallel to the first axis between the one or moreplants and the sampling system. The plug handler is further configuredto identify the individual plant of the one or more plants and securethe individual plant for movement relative to the one or more plants.The system also includes a tray table mechanically coupled to the frameassembly. The tray table is configured to collectively receive the oneor more plants and move relative to the frame assembly parallel to thesecond axis. The system also includes a popper mechanically coupled tothe frame assembly and configured to move relative to the frame assemblyparallel to the first axis and the third axis. The tray table receivesthe one or more plants and the controls system operates to move the traytable, the popper, and the plug handler to move each device proximate tothe individual plant of the one or more plants. The popper engages theindividual plant to move the plant along the third axis into the plughandler, and the plug handler secures the individual plant and movesalong the first axis to transport the individual plant to the samplingsystem.

In some embodiments, the tray table is coupled to a medial portion ofthe frame assembly; the plug handler is coupled to an upper portion ofthe frame assembly, the upper portion being above the medial portion;and the popper is coupled to a lower portion of the frame assembly, thelower portion being below the medial portion. The tray table may have aplurality of openings. Each opening contains a respective plant of theone or more plants, and the popper is configured to engage each plantfrom below the tray table through each respective opening of theplurality of openings.

In some embodiments, the plug handler includes a plug handler headcoupled to an end of the plug handler proximate to the tray table, and agripper mounted to the plug handler head and configured to selectivelygrip the individual plant. The plug handler head may also include anidentification device coupled to the plug handler head and configured toidentify the individual plant when the plug handler head is proximate tothe individual plant.

In some embodiments, each plant of the one or more plants is labeledwith a bar code and the plant identification device is a bar codescanner configured to scan the bar code positioned on each individualplant of the one or more plants.

In some embodiments, each plant of the one or more plants is labeledwith a radio frequency identification (RFID) tag and the plantidentification device is an RFID scanner configured to scan the RFID tagpositioned on each individual plant of the one or more plants.

In some embodiments, the gripper is a first gripper and the plug handlerfurther includes the first gripper extending outwardly from the plughandler head in a first direction parallel to the first axis, and asecond gripper coupled to the plug handler head and extending outwardlyfrom the plug handler head in a second direction parallel to the firstaxis and opposite the first direction. The identification device ispositioned between the first gripper and the second gripper, and thefirst gripper and the second gripper are configured to grip and holdseparate plants of the one or more plants. The plug handler head isrotatable so that the first gripper and the second gripper selectivelyoccupy opposite locations on the first axis.

In some embodiments, the sampling system includes a plant positionercoupled to the frame assembly and configured to receive the individualplant from the plant handling system, and a chuck coupled to the frameassembly proximate to the plant positioner. The chuck is configured tosecure a portion of the individual plant for sampling. The samplingsystem also includes an imager coupled to the frame assembly proximateto the chuck. The imager is configured to determine one or more of thecolor, shape, and size of the individual plant. The plant positioner isfurther configured to manipulate the individual plant to allow theimager to determine one or more of the color, shape, and size of theindividual plant and determine the sampling location in response. Theplant positioner positions the individual plant to align the samplinglocation with the chuck. The sampling system also includes a samplercoupled to the frame assembly proximate to the chuck. The sampler isconfigured to remove a tissue sample from the sampling location. Thesampling system still further includes a sample transport system coupledto the frame assembly proximate to the chuck that is configured toretrieve the tissue sample of the individual plant from the chuck andtransport the sample portion to the storage system. The controls systemis communicatively coupled to the plant positioner, the chuck, theimager, the sampler, and the sample transport system to control andoperate the plant positioner to receive the individual plant from theplant handling system, manipulate the individual plant foridentification by the imager, and position the plant so that the samplelocation is proximate to the chuck. The controls system is furtherconfigured to operate the chuck to secure the sample location to thechuck, to operate the sampler to remove the tissue sample from thesampling location, and to operate the sample transport system totransport the tissue sample to the storage system.

In some embodiments, the plant positioner is a motorized rotary gripperconfigured to grip and rotate the individual plant.

In some embodiments, the plant positioner moves parallel to the secondaxis.

In some embodiments, the chuck is a vacuum chuck configured to supply anair pressure at a location proximate to a surface of the chuck, whereinwhen the plant positioner brings the sampling location proximate to thechuck, the pressure draws the sample location to the chuck.

In some embodiments, the pressure flattens a portion of the samplelocation.

In some embodiments, the pressure is a pressure less than an ambientpressure at the automatic plant sampling system.

In some embodiments, the pressure is a pressure greater than an ambientpressure at the automatic plant sampling system.

In some embodiments, the imager is an imaging system having at least onecamera.

In some embodiments, the sampler is a laser cutting device operable inresponse to the image of the individual plant generated by the imagingsystem.

In some embodiments, the sampler is a laser cutting device.

In some embodiments, the sampler is a device having a plurality ofknives configured to cut the tissue sample from the sample location.

In some embodiments, the sampler is a tissue squeezing device configuredto apply compressive pressure to remove the tissue sample from thesample location.

In some embodiments, the tissue squeezing device is a hole punchingdevice.

In some embodiments, the sample transport system applies an air pressureless than the ambient pressure of the automatic plant sampling system todraw the tissue sample to a carrier for transportation to the storagesystem.

In some embodiments, the storage system includes a microtiter having aplurality of isolated locations. Each location has a scannable labelassociated with the individual plant. The storage system also includes acooling system positioned proximate to the microtiter and configured tomaintain the microtiter at a pre-determined temperature.

In some embodiments, the cooling system is dry ice disposed adjacent toan exterior of the microtiter.

In yet another illustrative embodiment, an apparatus for selecting,sampling, and storing a tissue sample from an individual plant of agroup of one or more plants is described. The apparatus includes a frameassembly having a first axis, a second axis orthogonal to the firstaxis, and a third axis orthogonal to the first axis and the second axis.The apparatus also includes a plant handler mounted to the frameassembly that has a plurality of grippers to manipulate the individualplant of the one or more plants to collect a tissue sample of theindividual plant. The apparatus also includes a sampler mounted to theframe assembly that has a sample identifier and a tissue samplecollection device to identify, collect, and store the tissue sample ofthe individual plant. The plant handler is configured to orient theplant within the sampler. The apparatus also includes a controls systemcommunicatively coupled to the plant handler and the sampler. Thecontrols system is configured to coordinate movement and operation ofthe plant handler and the sampler.

In some embodiments, the plant handler includes a plug handler havingone or more of the plurality of grippers. The plug handler ismechanically coupled to the frame assembly and is configured to moveparallel to the first axis between the one or more plants and thesampler. The plug handler is further configured to identify theindividual plant of the one or more plants and grip the individual plantwith one or more of the plurality of grippers for movement of theindividual plant relative to the one or more plants. The plant handlerincludes a tray table mechanically coupled to the frame assembly. Thetray table is configured to collectively receive the one or more plantsand move relative to the frame assembly parallel to the second axis. Theplant handler also includes a popper mechanically coupled to the frameassembly and configured to move relative to the frame assembly parallelto the first axis and the third axis. The tray table receives the one ormore plants and the controls system operates to move the tray table, thepopper, and the plug handler proximate to the individual plant of theone or more plants. The popper engages the individual plant to move theplant along the third axis into a gripper of the plurality of grippers,which grips the individual plant and moves along the first axis totransport the individual plant to the sampler.

In some embodiments, the plug handler also includes a plug handler headcoupled to an end of the plug handler proximate to the tray table. Theone or more grippers are mounted to the plug handler head, and the plughandler head is rotatable to selectively position each gripper of theplurality of grippers in a plan containing the first axis. The plughandler also includes an identification device coupled to the plughandler head and configured to identify the individual plant.

In some embodiments, each plant of the one or more plants is labeledwith a bar code and the plant identification device is a bar codescanner configured to scan the bar code positioned on each individualplant of the one or more plants.

In some embodiments, each plant of the one or more plants is labeledwith a radio frequency identification (RFID) tag and the plantidentification device is an RFID scanner configured to scan the RFID tagpositioned on each individual plant of the one or more plants.

In some embodiments, the apparatus further includes a plant positionerhaving a motorized rotary gripper coupled to the frame assembly. Theplant positioner is configured to move parallel to the second axis andreceive the individual plant from the plant handler. The apparatus alsoincludes a chuck coupled to the frame assembly proximate to the plantpositioner. The chuck is configured to engage a portion of theindividual plant for sampling. The sampler includes an imager coupled tothe frame assembly proximate to the chuck. The imager is configured todetermine one or more of the color, shape, and size of the individualplant. The sampler also includes a cutter coupled to the frame assemblyproximate to the chuck. The cutter is configured to remove a tissuesample from the sampling location of the individual plant. The sampleralso includes a tissue sample transporter (TST) coupled to the frameassembly proximate to the chuck that is configured to retrieve thetissue sample of the individual plant from the chuck and transport thetissue sample to a storage assembly. The plant positioner is furtherconfigured to manipulate the individual plant to allow the imager todetermine one or more of the color, shape, and size of the individualplant and determine the sampling location. The plant positionerpositions the individual plant to align the sampling location with thechuck. The controls system is communicatively coupled to the plantpositioner, the chuck, the imager, the cutter, and the tissue sampletransporter to control and operate the plant positioner to manipulatethe individual plant for identification by the imager and position theplant so that the sample location is proximate to the chuck. The controlsystem is further configured to operate the chuck to secure the samplelocation to the chuck, operate the sampler to remove the tissue samplefrom the sampling location, and operate the tissue sample transporter totransport the tissue sample to the storage assembly.

In some embodiments, the chuck is a vacuum chuck configured to supply apressure less than an ambient pressure at a location proximate to asurface of the chuck. When the plant positioner brings the samplinglocation proximate to the chuck, the pressure draws the sample locationto the chuck.

In some embodiments, the pressure flattens a portion of the samplelocation.

In some embodiments, the imager is an imaging system having at least onecamera.

In some embodiments, the sampler is a laser cutting device.

In some embodiments, the sampler is a device having a plurality ofknives configured to cut the tissue sample from the sample location.

In some embodiments, the sampler is a hole punching device configured toapply compressive pressure to remove the tissue sample from the samplelocation.

In some embodiments, the collection vessel includes a microtiter havinga plurality of isolated locations each location having a scannable labelassociated with the individual plant, and a dry ice receptacle forplacing dry ice proximate to the microtiter to maintain the microtiterat a pre-determined temperature.

In another illustrative embodiment, a method for automatically samplingindividual plants from a group of one or more plants is described. Themethod loads flats containing a plurality of plants into an automaticsampling device having a plant handling system and a plant samplingsystem. Each plant has a unique identifier. The method selects anindividual plant from the tray and secures the individual plant in theplant handling system. The method moves the individual plant with theplant handling system to the plant sampling system, and secures theindividual plant with the plant sampling system. The method releases theindividual plant with the plant handling system. The method identifiesat least one of a color, size, and shape of the individual plant withthe plant sampling system, and determines a sampling location of theindividual plant based on at least one of the color, size, and shape ofthe individual plant. The method secures the individual plant forsampling with the plant sampling system, and isolates a tissue samplefrom the individual plant with the plant sampling system. The methodtransports the tissue sample to a collection vessel with the plantsampling system and associates the tissue sample with the identifier ofthe individual plant. The method stores the tissue sample for testingand returns the individual plant for further cultivation with the planthandling system.

In some embodiments, the method engages the individual plant with apopper of the plant handling system to move the individual plant from atray table of the plant handling system into a gripper of the planthandling system.

In some embodiments, the method images the individual plant with animaging system of the plant sampling system.

In some embodiments, to the determine shape, size, and color of theindividual plant, the method selects the sampling location based on apredetermined size, color, and shape that corresponds with desiredcharacteristics.

In some embodiments, the desired characteristics include a size, color,and shape associated with an increased likelihood of plant survival.

In some embodiments, the method orients the individual plant so that thesample location is proximate to a chuck of the plant sampling system.The method positions the sample location with the chuck so that thesample location is disposed for isolating, and inhibits movement of thesample location with the chuck.

In some embodiments, the method applies a pressure lower than an ambientpressure of the plant sampling system to draw the sample location to thechuck.

In some embodiments, the method cuts the sample location with a lasercutting apparatus to remove the tissue sample from the individual plant.

In some embodiments, the method collects the tissue sample with a vacuumsystem and carries the tissue sample to a tissue sample collection plateassociated with the individual plant.

In some embodiments, the method repeats until each individual plant ofthe plurality of plants is sampled.

BRIEF DESCRIPTION OF THE DRAWINGS

Illustrative embodiments are described in detail below with reference tothe attached drawing figures, which are incorporated by referenceherein, and wherein:

FIG. 1 illustrates a schematic diagram of an illustrative system forautomatic plant tissue sampling;

FIG. 2 illustrates a schematic perspective view of an illustrativeautomatic plant tissue sampler;

FIG. 3 illustrates a schematic detail view of an illustrative popper ofthe automatic plant tissue sampler of FIG. 2 ;

FIG. 4 illustrates a schematic rear elevation view of an illustrativeplant sampling portion of the automatic plant tissue sampler of FIG. 2 ;

FIG. 5 illustrates a schematic perspective view of the plant samplingportion of the automatic plant tissue sampler of FIG. 2 ;

FIG. 6 illustrates a flowchart demonstrating an illustrative process ofautomatically sampling plant tissue;

FIG. 7 illustrates a flowchart demonstrating an illustrative process ofautomatically moving plants for sampling plant tissue;

FIG. 8 illustrates a schematic diagram representing exemplary componentsof an illustrative controls system for automatically sampling planttissue in accordance with the principles of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The invention allows for the automated and high-throughput sampling ofplant tissue, representing a significant advance over the prior art. Theprocesses provided herein increase the speed of tissue sampling whiledecreasing the error rate associated with prior art processes thatrelied on manual sampling. In addition, the processes provided hereinallow for tissue sampling of plants at an earlier post-germination stageof plant cultivation. Still further, the processes provided herein mayfully automate the tissue sampling process while leaving each sampledplant within a growth medium, allowing continued cultivation of theplant following tissue sampling. For example, the system provided hereinmay sample a high volume of samples in a short time while improvingconsistency of the samples and accuracy of association of the sampleswith the plant from which the sample was selected. The process asprovided may be applied to sampling of tissues from any plant species toprovide tissue samples for analytical testing that could involve DNA,RNA, protein, or any other analytical tests. The disclosed embodimentscan be used on transgenic tissue culture regenerants (R0 plants) orsubsequent generations of transgenic plants (R1 and beyond). Thedisclosed embodiments can also be used to collect samples fromnon-transgenic plants, for example, for the purposes of performingmolecular marker analysis in the context of molecular breeding programs.Other uses could include testing for metabolites in chemical screens orphysiological assays. The exemplary embodiments also provide a flexibleprocess that incorporates input and output. The exemplary embodimentsalso provide, in certain aspects, for delivering plant tissues forsampling. In one embodiment, this includes vacuum-assisted positioningand flattening of leaf or other plant tissues to facilitate sampling.Still further, the exemplary embodiments provide a non-contact lasercutting system that reduces cross contamination of tissue samples. Inaddition, the disclosed embodiments may accommodate a larger range ofplant sizes. The disclosed embodiments also provide repeatable samplesize and quality of tissue samples through the use of automaticallycontrolled imaging and cutting of plant tissue. In a exemplaryembodiment, the process may select and process each tissue sample in aslittle as 10 seconds.

FIG. 1 schematically illustrates an illustrative automatic plant tissuesampling system (APTSS) 11. As shown in FIG. 1 , the illustrative APTSS11 includes, in one embodiment, a plant handler 13, an imager 14, aplant sampler 15, a collection vessel 17, and automatic controls 19.Generally, the plant handler 13 may be a mechanical assembly having onemore apparatuses configured to transport a plant to the sampler 15 froma group of plants placed proximate to the APTSS 11. The imager 14includes one or more apparatuses configured to identify a suitablesampling location of the plant. The sampler 15 includes, for example,one or more components configured to take a tissue sample from thesampling location of the plant and transport the tissue sample to thecollection vessel 17. In one exemplary embodiment, the sampler 15includes a vacuum chuck, described in more detail below, configured tosecure the plant for tissue sampling without damaging the plant. Thecollection vessel 17, for example, includes one or more componentsconfigured to store the tissue samples in a manner that allows fortesting of the tissue samples at a desired time. The automatic controls19 include, in one embodiment, suitable control equipment configured tooperate the plant handler 13, the imager 14, the sampler 15, and thecollection vessel 17. In the illustrated embodiment, the automaticcontrols 19, which may include one or more computer processors andcommunication electronics, are communicatively coupled to the planthandler 13, the imager 14, the sampler 15, and the collection vessel 17for transmission of operative signals between the automatic controls 19and the plant handler 13, the imager 14, the sampler 15, and thecollection vessel 17 for operation thereof.

A plurality of plants may be brought to the APTSS 11 at a general entrylocation 21, a location of the APTSS 11 where the plant handler 13 mayaccess the plurality of plants. In some exemplary embodiments, theplurality of plants are conveyed to the entry location 21 of the APTSS11 by an operator or other laborer. The operator or laborer may, forexample, assemble the plurality of plants onto a flat or other memberand physically convey the plants to the entry location 21 of the APTSS11. As used herein, “flat” refers to any object or device suitable forholding, supporting, arranging or otherwise carrying or moving one ormore given sample(s), such as plants or other sources of tissue to besampled in accordance with the invention.

In other exemplary embodiments, a plurality of plants are conveyed tothe APTSS 11 by an automatic conveyance system 27 that may deposit theplurality of plants with the APTSS 11 in any suitable manner. In anon-limiting example, the conveyance system 27 may receive the pluralityof plants at a separate location and convey the plurality of plantsalong a conveyor, such as a conveyor belt or any other suitablemotorized apparatus. The plants may be hand-assembled by an operator andplaced onto the conveyor assembly. In other embodiments, the plants maybe automatically assembled and conveyed on the conveyance system 27. Asused herein, assembly of the plants refers to the grouping of more thanone plant into a batch or flat to be processed by the APTSS 11.

The plant handler 13, in one embodiment, selects a plant from theplurality of plants at the entry location 21 and moves the plant fromthe plurality of plants to the imager 14. In an exemplary embodiment,the plant handler 13 includes an overhead gantry apparatus having amechanical gripping device configured to pick up a plant from theplurality of plants and move the plant to the imager 14. In anotherexemplary embodiment, the plant handler 13 may include an apparatusconfigured to place a plant on a conveyor that may move the plant to theimager 14. Alternative configurations of the plant handler 13 to movethe plant to the imager 14 may be utilized. There, the imager 14 imagesthe plant and identifies a suitable sampling location. The imager 14 mayalso or alternatively include an ultraviolet camera, an infrared camera,or any other suitable imaging device. The imager 14 may utilize one ormore components, operate in one or more frequency spectrums, and haveresolution that provides accuracy in accordance with the principles ofthe present invention. For example, the imager 14 may include a visiblelight camera configured to image the plant to determine the suitablesampling location. In one exemplary embodiment, the plant may be a cornplant, or any other species, and the imager 14 may image the leaf. Theautomatic controls 19 may then determine that the leaf is an appropriatesize or shape, for example, larger than 6 mm by 12 mm rectangle, have asuitable hue, for example, have a green hue, be of a suitable shape, forexample, substantially rectangular, or some combination thereof. Theplant sampler 15 then removes a tissue sample from the sampling locationand transports the tissue sample to, for example, the collection vessel17. In one exemplary embodiment, the plant sampler 15 may be a lasercutting device, a mechanical apparatus having a plurality of knives, ahole-punching device, or any other device configured to remove a tissuesample from a plant without destroying the plant. A person skilled inthe art will understand that the imager 14 and the sampler 15 may beseparate devices or may be a single device performing the functions ofboth. The collection vessel 17 may be any suitable storage deviceconfigured to isolate each tissue sample from other tissue samples andmaintain the tissue samples in a suitable condition for testing.

In an exemplary embodiment, the plant handler 13 retrieves the sampledplants from the sampler 15 and transports the sampled plants to asampled plant location 23. In some embodiments, the sampled plantlocation 23 may be the entry location 21. In other exemplaryembodiments, the imager 14 may determine that the plant may not besuitable for sampling. For example, the imager 14 may communicate withthe automatic controls 19 to compare an image of the plant with imagesof plants suitable for testing. For example, the automatic controls 19may identify a plant leaf of an imaged plant to have a green hue that isassociated with plants that are not healthy. In another embodiment, theautomatic controls 19 may identify that an imaged leaf may have a sizethat is too small for sampling, for example, less than 12 mm×6 mmrectangle. In still another embodiment, the automatic controls 19 mayidentify that the leaf shape may not be sufficiently rectangular to besuitable for sampling. In response to determining that the plant isunsuitable for sampling, the plant handler 13 may transport the plantfrom the sampler 15 to an unsuitable plant location 25. In an exemplaryembodiment, plants that are not suitable for sampling may be too smallor may not yet have reached suitable maturity. These plants may beallowed to further mature or grow before being sampled by the APTSS 11.A person skilled in the art will understand that the sampled plantlocation 23 and the unsuitable plant location 25 may be the entrylocation 21. A person skilled in the art will also understand that theautomatic controls 19 may include suitable mechanisms, includingsoftware, controllers, and the like, configured to identify individualplants relative to the plurality of plants, identify predeterminedproperties of those plants, and store that information so that theplurality of plants may be maintained in a single location, such as atthe entry location 21, of the APTSS 11.

In some embodiments, the plant handler 13, the sampler 15, and thecollection vessel 17 are coupled to a frame assembly 29. The frameassembly 29 may be any suitable assembly configured to support the planthandler 13, the imager 14, the sampler 15, and the collection vessel 17for operation thereof. In an exemplary embodiment, the frame assembly 29may include a support structure formed of individual members coupledtogether, one or more components of a location where the APTSS 11 ishoused, such as a building floor, foundation, overhead beam, or anysuitable support structure.

In the embodiments described herein, the plant or the plurality ofplants being sampled may be any suitable plant, for example, corn,soybean, cotton, canola, alfalfa, wheat, sugarcane, rice, or the like.The tissue samples may be taken from any desired location of the plant,for example, from a seed, root, stem, inner stem, stalk or leaf, or thelike. For descriptive purposes, a plant of the plurality of plantsherein includes any plant parts or plant tissues, which includes tissuecultures. The plants and tissues may or may not be comprised in agrowing media. The growing medium may be any suitable substance in whichplants or plant tissues may grow or be maintained. A person skilled inthe art will understand that the disclosed embodiments include otherplants and parts configured in other arrangements.

As shown in FIG. 2 , the frame assembly 29 may have a plurality of beamsor members coupled together to provide a working frame for the APTSS 11.While the frame assembly 29 is illustrated as a box frame assembly of aplurality of members having a plurality of openings, a person skilled inthe art will understand that the frame assembly may be any suitablesupportive apparatus or group of apparatuses such that the frameassembly may support the plant handler 13, the imager 14, the sampler15, and the collection vessel 17 relative to one another for operationof the APTSS 11. In the illustrated embodiment, the frame assembly 29includes a lower portion 31, a medial portion 33, and an upper portion35. Lower portion 31, medial portion 33, and upper portion 35 areincluded herein for reference only and are not intended to limit theembodiments of the frame assembly 29 or the APTSS 11. The frame assembly29 may have three orthogonal axes, an x-axis 37, a y-axis 39, and az-axis 41. These axes are presented for descriptive purposes to aid inillustrating the relationships between components of the APTSS 11 andnot to limit the embodiments herein. A person skilled in the art willunderstand that any suitable coordinate reference system may be used torelate the components of the APTSS 11 based on the particularapplication of the APTSS 11.

Continuing to refer to FIG. 2 , the plant handler 13 includes, in oneembodiment, a popper assembly or popper 43, a plant receiving area ortray table 45, and an overhead gantry assembly or gantry 47. Popper 37may be a device movably coupled to the frame assembly 29 in the lowerportion 31 of the frame assembly 29 and may be configured to moveparallel to the x-axis 37. Referring to FIG. 3 , the popper 43 includes,in one embodiment, an actuable member 49 configured to move at least aportion of the popper 43 parallel to the z-axis 41. In the illustratedembodiment, the actuable member 49 may be a pneumatically,hydraulically, mechanically, or electrically operable cylinderconfigured to move a rod portion 51 parallel to the z-axis 41 toward themedial portion 33 of the frame assembly 29. The automatic controls 19may be communicatively coupled to the popper 43 to control movement ofthe popper 43 parallel to the x-axis 37 and control operation of the rodportion 51, i.e. the application of pneumatic, hydraulic, or electricpower, to move the rod portion 51 toward the medial portion 33 of theframe assembly 29. In an exemplary embodiment, the rod portion 51includes an insertive device configured to be inserted into a lowerportion of a plant of the plurality of plants to prevent relative motionbetween the plant and the rod portion 51 when the rod portion 51 engagesthe plant. A person skilled in the art will recognize that the popper 43may include one or more controllers, motors, and the like configured tostart and stop motion of the popper 43 parallel to the x-axis 37 and thez-axis 41 in response to communicative inputs from, for example, theautomatic controls 19. In one exemplary embodiment, the popper 43 maytranslate about 200 mm parallel to the x-axis 37 and the rod portion 51may move parallel to the z-axis 41 about 50 mm.

Referring again to FIG. 2 , the tray table 45 may be movably coupled tothe medial portion 33 of the frame assembly 29 so that the tray table 45may be generally over the popper 43. The tray table 45 is configured, inone embodiment, to receive the plurality of plants for sampling by theAPTSS 11. In the illustrated embodiment, the tray table 45 includes oneor more openings configured to receive flats having the plurality ofplants. As shown in FIG. 2 , the tray table 45 has three openings eachof a suitable size and shape to allow placement of a flat 46 in therespective opening. An alternative number of openings in the tray table45 may be utilized. Each flat 46 has a plurality of openings configuredto receive a plant. In the illustrated embodiment, each opening in theflat 46 passes entirely through the flat 46 parallel to the z-axis 41and accommodates a plug or other growth medium matrix (soil, artificialsoil, rockwool, etc.) within each opening. In an exemplary embodiment,the tray table inhibits movement of the flats 46 relative to the traytable 45 along the x-axis 37, the y-axis 39, and the z-axis 41 duringoperation of the APTSS 11 to ensure that the plants are in a locationknown by the automatic controls 19 and reachable by the gantry 47.

In the illustrated embodiment, the tray table 45 may receive three flats46, a first flat 53, a second flat 55, and a third flat 57. The firstflat 53 may be a flat 46 containing the plurality of plants to besampled. The first flat 53 may be delivered by an automatic conveyancesystem, an operator or laborer, or the like. The first flat 53 may bedeposited or placed in one of the openings of the tray table 45. Thesecond flat 55 may be placed in a second opening of the three openingsof the tray table 45 and contain no plants when the sampling processbegins. In the illustrated embodiment, the second flat 55 may be used tostore a plant after the plant has been sampled. The third flat 57 may beplaced in a third opening of the tray table 45 and also include noplants when the sampling process begins. In the illustrated embodiment,the third flat 57 may serve as an isolation flat where plants that arenot suitable for testing, i.e., the plant may be too small, may beplaced for further growth or other treatment. As shown, the first flat53 may be placed at a first end of the tray table 45 followed by thesecond flat 55 and the third flat 57 at a second end of the tray table45. A person skilled in the art will recognize that the flats 53, 55, 57may be placed in any order provided the placement of each flat 53, 55,57 is known by the automatic controls 19, and the flat containing theplurality of plants to be sampled is accessible to the popper 43. Insome embodiments, each opening of the tray table 45 may receive a firstflat 53 to increase the speed of tissue sampling by reducing theinterval between plant groups. In these embodiments, the plants may besampled and returned to their original location in the first flat 53from which they were selected for sampling. The tray table 45 may becoupled to a plurality of controllers, motors, and the like, andcommunicatively coupled to the automatic controls 19 so that the traytable 45 may be moved parallel to the y-axis 39.

As shown in FIG. 2 , the gantry 47 is movably coupled to the upperportion 35 of the frame assembly 29 so that at least a portion of thegantry 47 is disposed over the tray table 45. In the illustratedembodiment, the gantry 47 includes a plug handler 59 having a plughandler head 61, a first gripper 63, and a second gripper 65. A personskilled in the art will understand that embodiments of the plant handler13 may have only the first gripper 63. Generally, the gantry 47 isconfigured to move the plug handler 59 parallel to the x-axis 37 andincludes suitable control mechanisms, control wiring, controllers,motors, and the like to accomplish motion. In addition, the automaticcontrols 19 may be communicatively coupled to the gantry 47 to operatethe control mechanisms for operation thereof. The plug handler 59 has afirst end proximate to the upper portion 35 of the frame assembly 29 anda second end proximate to the tray table 45. The plug handler head 61 iscoupled to the second end of the plug handler 59. In the illustratedembodiment, the plug handler head 61 may be rotatable so that the firstgripper 63, and the second gripper 65 may rotate about an axis passingthrough the plug handler 59 parallel to the z-axis 41. The grippers 63,65 may be a suitable device configured to grip a plant of the pluralityof plants when the plant of the plurality of plants is placed proximateto the grippers 63, 65 by the popper 43. In the illustrated embodiment,the grippers 63, 65 are clamping devices that may be pneumatically,hydraulically, electrically or otherwise actuated to apply a compressiveforce to the plug of the plant when the plug is placed proximate to thegrippers 63, 65.

In an operative embodiment, the APTSS 11 may receive the first flat 53from any suitable conveyance system 29 (FIG. 1 ). The automatic controls19 may operate to move the popper 43, the tray table 45, and the gantry47 so that the popper 43 is positioned beneath one of the plants of thefirst flat 53 and the first gripper 63 is positioned above the sameplant. The automatic controls 19 may then actuate the actuable member 49to drive the rod portion 51 upwards toward the plant, engaging theinsertive device with the plant. The rod portion 51 may have asufficient throw to drive the plant from the first flat 53 upwards andadjacent to the first gripper 63. There, the first gripper 63 may beactuated by the automatic controls 19 to close on the plant and securethe plant within the first gripper 63. The rod portion 51 may beretrieved by the actuable member 49, leaving the plant at the elevationof the first gripper 63. In some embodiments, the plug handler 59 may bemovable parallel to the z-axis 41 so that the plug handler may belowered proximate to the tray table 45 to retrieve the plant without theassistance of the popper 43. Once the plant is secured in the firstgripper 63, the gantry 47 may be actuated by the automatic controls 19to move parallel to the x-axis 37 to bring the plug handler 59 and thefirst gripper 63 proximate to the imager 14 and the sampler 15. A personskilled in the art will recognize that the plant handler 13 may includeone or more controllers, motors, and the like configured to start andstop motion of the tray table 45, the gantry 47, the plug handler 59,the first gripper 63, and the second gripper 65 parallel to the x-axis37, the y-axis 39, and the z-axis 41 in response to communicative inputsfrom, for example, the automatic controls 19.

In an exemplary embodiment, the plug handler head 61 includes a readeror scanner 67, such as a bar code scanner or camera. The scanner 67 maybe any suitable device configured to identify a code, label, tag,object, or other identifier placed on, in proximity to, or incorporatedinto an individual plant of the plurality of plants. Alternatively, thescanner 67 may be configured to recognize an image or shape of a plantor carrier of the plant. In illustrative embodiments the code or labelmay be a bar code, a radio frequency identification tag, or any othersuitable identifier. The scanner 67 may be communicatively coupled tothe automatic controls 19. In the illustrated embodiment, the scanner 67is positioned so that the a bar code disposed on each flat 46 may beread by the scanner 67. The scanner 67 may communicate thisidentification information to the automatic controls 19, and theautomatic controls 19 may store the identification information of thebar code in an internal or external memory of the automatic controls 19.The size, shape, and number of plants on each flat 46 is also stored inthe automatic controls 19. The automatic controls 19 may track eachplant based on the plant's position on the flat 46 associated with thescanned bar code for the flat 46. In another exemplary embodiment, thescanner 67 may be coupled to the plug handler head 61 so that a bar codeon the plant may be read by the scanner 67 when the plant is secured inthe first gripper 63. In yet another exemplary embodiment, each plantmay be separately conveyed to the APTSS 11 without a flat 46. Each plantmay then also include a bar code or other identifier that may be read bythe scanner 67.

In one exemplary embodiment, the identifier associated with each plantmay include information relating to the plant type, planting date,germination date, variety, or attribute information. This attributeinformation could include transgenes conferring resistance to heat,cold, fungi, parasitcs, viruses, bacteria, drought, and the like. Theattribute information may also include size, shape, or color at previoussamplings. This information may be in a database stored in theidentifier or in a separate location in communication with theidentifier or scanner 67 so that the information may be accessed by theautomatic controls 19 or any other suitable device. In addition, theimager 14 may be in communication with the database so that informationrelating to the size shape and color of the sampled plant prior tosampling may also be stored and associated with the identifier. A personskilled in the art will understand that the database may include anyother suitable information as needed for the particular application ofAPTSS 11.

The gantry 47 is generally disposed over the entry location 21 and thetray table 45, and includes a portion proximate to the imager 14 and thesampler 15. The gantry 47 may move the plug handler 59 proximate to theimager 14 and the sampler 15, where the plant may be transferred fromthe first gripper 63 to the sampler 15 as described in more detailbelow. In an exemplary embodiment, following sampling of the plant, theplant may be transferred back to the plug handler 59 and transported tothe second flat 55.

Continuing to refer to FIG. 2 , the sampler 15 includes a plantpositioner 69, a chuck 71, a cutter 75, and a tissue sample transporter(TST) 81. As shown, the imager 14 is proximate to the sampler 15 so thatthe sampler 15 may be guided by the imager 14 to remove a tissue sampleas described in more detail below. As shown in FIG. 5 , the plantpositioner 69 includes a plant gripper 77. Plant gripper 77 is similarto the first and second grippers 63, 65 of the plant handler 13 and mayoperate in a similar manner in response to control inputs from theautomatic controls 19. In addition, the plant gripper 77 may berotatable about an axis parallel to the x-axis 37 so that the plantgripped by the plant gripper 77 may be rotated about an axis parallel tothe x-axis 37. In addition, the plant positioner 69 is movably coupledto the frame assembly 29 so that the plant positioner 69 may moveparallel to the y-axis 39. Rotation of the plant about the axis parallelto the x-axis 37 and translation of the plant parallel to the y-axis 39permits the plant to be oriented for proper imaging by the imager 14 andsampling by the sampler 15 as described in more detail below. A personskilled in the art will recognize that the plant positioner 69 mayinclude suitable control mechanisms to operate the plant positioner 69as disclosed herein. A person skilled in the art will recognize that theplant positioner 69, the chuck 71, the cutter 75, and the TST 81, mayinclude one or more controllers, motors, and the like configured tostart and stop motion parallel to the x-axis 37, the y-axis 39, and thez-axis 41 in response to communicative inputs from, for example, theautomatic controls 19.

Referring to FIG. 4 , a partial side elevation view of the imager 14 andthe sampler 15 is shown. As shown in FIG. 4 , the chuck 71 may bepositioned on the medial portion 33 of the frame assembly 29. The cutter75 may be coupled to the upper portion 35 of the frame assembly 29. Inthe illustrated embodiment, the cutter 75 may be a laser cutterconfigured to direct a cutting laser at a surface of the chuck 71. Inother embodiments, the cutter 75 may be a plurality of knives configuredto cut the tissue sample, a tissue squeezing device configured to applycompressive pressure to remove the tissue sample, or a hole punchingdevice. In the illustrated embodiment, the chuck 71 may be a vacuumchuck supplied with a pressure source adapted to generate a pressureless than the ambient pressure of APTSS 11 above the surface of thechuck 71. The chuck 71 may have a porous platform having a fanpositioned relative to the porous platform so that, when operated, thefan may draw a high volume of air through the porous platform. Theimager 14 may also be coupled to the upper portion 35 of the frameassembly 29 so that the plant may be exposed to the imager 14 when theplant is proximate to the chuck 71. In the illustrated embodiment, theimager 14 may be an imaging system configured to image the plant anddetermine one or more of the plant size, shape, or color. The imager 14may be a visible, infrared, ultraviolet, or any other suitable camera.The plug handler 59 may be brought proximate to the plant positioner 69and the plug handler head 61 rotated so that the plant may betransferred from the first gripper 63 to the plant positioner 69. In anembodiment, the plant gripper 77 and the first gripper 63 occupy a samehorizontal plane so that the transfer may occur by having the plantgripper 77 actuate and secure the plant and the first gripper 63 actuateto release the plant.

The plant positioner 69 may receive the plant from the first gripper 63and position the plant proximate to the chuck 71 so that the imager 14may image the plant. In response to the image generated by the plant,the automatic controls 19 may operate the plant positioner 69 toposition the sampling location adjacent the surface of the chuck 71. Inone exemplary embodiment, the plant positioner 69 orients the plant sothat a stem or stalk of the plant is at a forty-five degree angle withthe porous platform of the chuck 71. The plant positioner 69 drags or“paints” the plant across the chuck 71 until the imager 14 identifies atip of a last leaf of the plant. In the illustrated embodiment, theautomatic controls 19 include appropriate control mechanisms, software,hardware, etc., to allow the automatic controls 19 to detect theupper-most green section of a leaf. Once detected, the automaticcontrols 19 then stop movement of the plant positioner 69 so that thetip of the last leaf of the plant is proximate to the chuck 71. Thechuck 71, actuated to generate a pressure less than the ambientpressures of the APTSS 11 on the surface of the chuck 71 prior toplacement of the tip of the last leaf of the plant proximate to thechuck 71, draws the sampling location into contact with the chuck 71 andimmobilizes the sampling location. In an embodiment, the samplinglocation may be a plant leaf having a suitable size, color, and shapesuch that removing a portion of the leaf will not irreparably damage theleaf causing death of the leaf or the plant. Once the sampling locationis immobilized by the chuck 71, the laser cutter or cutter 75 may beoperated to cut a portion of the sampling location from the plant. In anexemplary embodiment, the sampler is guided by the image produced by theimager 14 to cut a tissue sample of a predetermined sized from thesampling location. In one exemplary embodiment, the tissue sample is arectangle having a length of about 6 mm and a width of about 12 mm. Aperson skilled in the art will recognize that the size of the tissuesample may vary as needed to accommodate the type of testing to beperformed with the tissue sample. A person skilled in the art will alsorecognize that the embodiments disclosed herein may be modified withoutchanging their general operation to accommodate a wide variety of tissuesample sizes.

Referring again to FIG. 5 , following cutting of the tissue sample fromthe sampling location, the automatic controls 19 may operate the TST 81to move to the chuck 71, retrieve the tissue sample, and transport thetissue sample to the collection vessel 17. In the illustratedembodiment, the TST 81 may be an assembly mounted to the upper portion35 of the frame assembly 29 proximate to the chuck 71. The TST 81 mayinclude a pneumatic pick and place device having a member 83 configuredto be positioned proximate to the surface of the chuck 71 and suppliedwith a vacuum pressure of sufficient strength to draw the tissue sampleoff of the surface of the chuck 71 when the vacuum pressure is appliedby the member 83. The member 83 may then be moved along the x-axis 37and the y-axis 39 to place the tissue sample proximate to the collectionvessel 17. A person skilled in the art will understand that TST 81includes suitable actuators, motors, and pneumatic system devicesconfigured to generate a vacuum pressure in member 83 and translatemember 83 relative to the chuck 71 and the collection vessel 17 to movethe tissue sample from the chuck 71 to the collection vessel 17. The TST81 and the sampler 15 may also include suitable devices configured toclean the TST 81 and the sampler 15 following collection of each tissuesample to prevent contamination between samples. The TST 81 can alsoinclude an additional bar code scanner to read barcodes associated withthe collection vessel 17.

As shown in FIG. 5 , the collection vessel 17 includes a storage bin 85positioned adjacent the sampler 15 so that the TST 81 may place themember 83 axially over the storage bin 85. The collection vessel 17 alsoincludes bodies 87 having a plurality of storage locations. In theillustrated embodiment, the bodies 87 may be microtiter plates eachhaving a plurality of wells or test tubes associated with a separateplant of the plurality of plants on the first flat 53. In an exemplaryembodiment, the TST 81 may bring the tissue sample proximate to thebodies 87 and release the vacuum pressure applied to the member 83 todeposit the tissue sample in a particular location or well of the bodies87, the particular location associated with the particular plant fromwhich the tissue sample was removed. In an exemplary embodiment, thestorage bin 85 is packed with dry ice to maintain the tissue samples ata predetermined temperature for storage thereof.

FIG. 6 illustrates a high-level flow chart that depicts logicaloperative steps of the APTSS 11 of FIGS. 1-5 , which may be implementedin accordance with an embodiment. As indicated at block 601, the processbeings by loading trays containing a plurality of plants into anautomatic sampling device. For example, the first flat 53 having theplurality of plants thereon is conveyed into the entry location 21 ofthe APTSS 11. Next, as indicated at block 603, the APTSS 11 selects anindividual plant and moves the individual plant to an imager. Forexample, the plant handler 13 selects and transports an individual planton the first flat 53 and transports the plant to the imager 14.

The APTSS 11 identifies at least one of the color, shape, or size of theplant at block 605. For example, the imager 14 of the sampler 15determines at least one of the plant's color, shape, or size. At block607, the APTSS 11 determines a sampling location in response to theidentification of at least one of the size, shape, and color of theplant. For example, the imager 14 determines the sampling location ofthe plant in response to previously identified at least one of the size,shape, and color of the plant. Next, as indicated at block 609, theAPTSS 11 secures the individual plant in the plant sampler. For example,the plant positioner 69 of the sampler 15 secures the plant within thesampler 15.

At block 611, the individual plant is secured and a tissue sample isisolated from the sampling location. For example, the plant positioner69 positions the sample location over the chuck 71. The chuck 71 mayimmobilize the sampling location of the plant with vacuum pressure. Thecutter 75 may cut the plant at the sampling location to create a tissuesample. Next, at block 613, the APTSS 11 transports and stores thetissue sample. For example, the TST 81 moves proximate to and appliesvacuum pressure to the tissue sample to secure the tissue sample to themember 83 of the TST 81. The TST 81 then transports the tissue sample tothe collection vessel 17, where the tissue sample is stored in anisolated location in collection vessel 17 associated with the plant fromwhich the sample was taken. The process ends at block 615, where theindividual plant is returned for further cultivation. For example, theplant positioner 69 transfers the plant to the plant handler 13, wherethe plant is transported to the second flat 55 and deposited in asuitable location therein.

Referring now to FIG. 7 , a high-level flow chart that depicts logicaloperative steps of the plant handler 13 performed by, for example, planthandler 13 of FIGS. 1-3 , which may be implemented in accordance with anembodiment. As indicated at block 701, the process begins by moving afirst gripper, gripper one, to the plant input flat. For example, thegantry 47 moves the plug handler 59 to position the first gripper 63proximate to the first flat 53. Next, as indicated at block 703, theplant handler 13 transfers an un-sampled plant from the input flat togripper one. For example, the plant handler 13 operates the popper 43 tomove the un-sampled plant from the first flat 53 into the first gripper63, and the first gripper 63 is actuated by the automatic controls 19 tosecure the un-sampled plant.

As indicated at block 705, the plant handler 13 moves a second gripper,gripper two, to the sample position. For example, the plant handler 13operates the gantry 47 to move the plug handler 59 proximate to theimager 14 and the sampler 15 so that the second gripper 65 is proximateto the plant positioner 69 of the sampler 15. Next, as indicated atblock 707, a sampled plant is transferred from a third gripper, gripperthree, to gripper two. For example, the sampled plant is transferredfrom the plant gripper 77 of the plant positioner 69 to the secondgripper 65. In an embodiment, the plant gripper 77 and the secondgripper 65 occupy a same horizontal plane so that the transfer may occurby having the second gripper 65 actuate and secure the sampled plant andthe plant gripper 77 actuate to release the sampled plant.

As indicated at block 709, gripper one is rotated to the plant sampleposition. For example, the plant handler 13 rotates the plug handlerhead 61 on an axis of the plug handler 59 parallel to the z-axis 41 toposition the first gripper 63 proximate to the plant gripper 77 of theplant positioner 69. Next, as indicated at block 711, the plant handler13 transfers the un-sampled plant from gripper one to gripper three. Forexample, the first gripper 63 of the plant handler 13 transfers theun-sampled plant from the first gripper 63 to the plant gripper 77 ofthe plant positioner 69. In an embodiment, the plant gripper 77 and thefirst gripper 63 occupy a same horizontal plane so that the transfer mayoccur by having the plant gripper 77 actuate and secure the un-sampledplant and the first gripper 63 actuate to release the un-sampled plant.

Referring to block 713, the plant handler 13 moves gripper two to theoutput flat. For example, the gantry 47 moves the plug handler 59 toposition the second gripper 65 proximate to the second flat 55. Asindicated at block 715, the plant handler 13 transfers the sampled plantfrom gripper two to that which then lowers the sampled plant onto thesecond flat 55.

Referring to FIG. 8 , an exemplary embodiment of a portion of theautomatic controls 19 is shown. The automatic controls 19 include a maincontroller 89 that may be communicatively coupled to the scanners 67,the cutter 75, the imager 14, a Monsanto LIMS, a plurality of integratedmotion drives 91, and a plurality of discreet input/output devices 93.The main controller 89 may be any suitable computing device or system,such as a programmable logic controller, a data processing system, orthe like, configured to receive input from the above listed devices andcommunicate with those same devices for operation thereof. A dataprocessing system suitable for storing and/or executing program codewill include at least one processor coupled directly or indirectly tomemory elements through a system bus. The memory elements can includelocal memory employed during actual execution of the program code, bulkstorage, and cache memories, which provide temporary storage of at leastsome program code in order to reduce the number of times code isretrieved from bulk storage during execution.

The plurality of integrated motion drives 91 are devices configured tomove a member parallel to one of the x-axis 37, the y-axis 39, thez-axis 41, a first axis of rotation, a second axis of rotation, or acombination thereof. For example, the plurality of integrated motiondrives 91 may be coupled to the elements of the plant handler 13 to movethe tray table 45, the popper 43, the gantry 47, the plant positioner69, and the TST 81. The integrated motion drives 91 are any suitabledevice configured to receive operative signals or instructions from themain controller 89 and translate or cause the translation of anassociated component of the APTSS 11 as described herein.

The discreet input/output devices 93 may be any suitable devices such aspneumatic sensors, temperature sensors, or the like configured tocommunicate signals to the main controller 89. Input/output or I/Odevices (including but not limited to keyboards, displays, pointingdevices, etc.) can be coupled to the system either directly or throughintervening I/O controllers. Network adapters may also be coupled to thesystem to enable the automatic controls 19 to become coupled to otherdata processing systems or remote printers or storage devices throughintervening private or public networks. Modems, cable modems, andEthernet cards are just a few of the currently available types ofnetwork adapters.

The disclosed embodiments may be used for collecting tissue samples frommost any plant species for analytical testing that could be DNA, RNA,protein, or any other analytical test. The disclosed embodiments may beused on transgenic tissue culture regenerants (R0 plants) or subsequentgenerations of transgenic plants (R1 and beyond). The disclosedembodiments may also be used to collect samples from non-transgenicplants, for example for the purpose of performing molecular markeranalyses in the context of molecular breeding programs, or for testingfor metabolites in chemical screens or physiology assays.

It is understood that the present invention may take many forms andembodiments. Accordingly, several variations may be made in theforegoing without departing from the spirit or scope of the invention.Having thus described the present invention by reference to certain ofits preferred embodiments, it is noted that the embodiments disclosedare illustrative rather than limiting in nature and that a wide range ofvariations, modifications, changes, and substitutions are contemplatedin the foregoing disclosure and, in some instances, some features of thepresent invention may be employed without a corresponding use of theother features. Many such variations and modifications may be consideredobvious and desirable by those skilled in the art based upon a review ofthe foregoing description of preferred embodiments. Accordingly, it isappropriate that the appended claims be construed broadly and in amanner consistent with the scope of the invention.

In the description and tables, a number of terms are used. In order toprovide a clear and consistent understanding of the specification andclaims, the following definitions are provided:

A: When used in conjunction with the word “comprising” or other openlanguage in the claims, the words “a” and “an” denote “one or more.”

Or: Unless otherwise indicated, as used throughout this document, “or”does not require mutual exclusivity.

Regeneration: The development of a plant from tissue culture.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence that has beenintroduced into the genome of a plant by transformation.

It should be apparent from the foregoing that an invention havingsignificant advantages has been provided. While the invention is shownin only a few of its forms, it is not just limited but is susceptible tovarious changes and modifications without departing from the spiritthereof.

What is claimed is:
 1. An apparatus for selecting, sampling, and storinga tissue sample from an individual plant of a group of one or moreplants, the apparatus comprising: a frame assembly having a first axis,a second axis orthogonal to the first axis, and a third axis orthogonalto the first axis and the second axis; a plant handler mounted to theframe assembly and having a plurality of grippers to manipulate theindividual plant of the one or more plants to collect a tissue sample ofthe individual plant; a sampler mounted to the frame assembly and havinga sample identifier and a tissue sample collection device to identify,collect, and store the tissue sample of the individual plant, the planthandler configured to orient the plant within the sampler; and acontrols system communicatively coupled to the plant handler and thesampler, the controls system configured to coordinate movement andoperation of the plant handler and the sampler.
 2. The automatic planthandling apparatus of claim 1, wherein the plant handler comprises: aplug handler having one or more of the plurality of grippers, the plughandler mechanically coupled to the frame assembly and configured tomove parallel to the first axis between the one or more plants and thesampler, the plug handler further configured to identify the individualplant of the one or more plants and grip the individual plant with oneor more of the plurality of grippers for movement of the individualplant relative to the one or more plants; a tray table mechanicallycoupled to the frame assembly, the tray table configured to collectivelyreceive the one or more plants and move relative the frame assemblyparallel to the second axis; a popper mechanically coupled to the frameassembly and configured to move relative to the frame assembly parallelto the first axis and the third axis; and wherein the tray tablereceives the one or more plants and the controls system operates to movethe tray table, the popper, and the plug handler proximate to theindividual plant of the one or more plants, the popper engages theindividual plant to move the plant along the third axis into a gripperof the plurality of grippers which grips the individual plant and movesalong the first axis to transport the individual plant to the sampler.3. The automatic plant handling apparatus of claim 2, wherein the plughandler further comprises: a plug handler head coupled to an end of theplug handler proximate to the tray table; the one or more grippersmounted to the plug handler head; the plug handler head rotatable toselectively position each gripper of the plurality of grippers in a plancontaining the first axis; and an identification device coupled to theplug handler head and configured to identify the individual plant,wherein: each plant of the one or more plants is labeled with a bar codeand the plant identification device comprises a bar code scannerconfigured to scan the bar code positioned on each individual plant ofthe one or more plants; or wherein each plant of the one or more plantsis labeled with a radio frequency identification (RFID) tag and theplant identification device comprises an RFID scanner configured to scanthe RFID tag positioned on each individual plant of the one or moreplants.
 4. The automatic plant sampling apparatus of claim 1, wherein:the plant handler further comprises: a plant positioner having amotorized rotary gripper coupled to the frame assembly and configured tomove parallel to the second axis and receive the individual plant fromthe plant handler; a chuck coupled to the frame assembly proximate tothe plant positioner, the chuck configured to engage a portion of theindividual plant for sampling; the sampler comprises: a imager coupledto the frame assembly proximate to the chuck, the imager configured todetermine one or more of the color, shape, and size of the individualplant; a cutter coupled to the frame assembly proximate to the chuck,the cutter configured to remove a tissue sample from the samplinglocation of the individual plant; a tissue sample transporter coupled tothe frame assembly proximate to the chuck and configured to retrieve thetissue sample of the individual plant from the chuck and transport thetissue sample to a storage assembly; the plant positioner furtherconfigured to manipulate the individual plant to allow the imager todetermine one or more of the color, shape, and size of the individualplant and determine the sampling location, the plant positionerpositioning the individual plant to align the sampling location with thechuck; and wherein the controls system is communicatively coupled to theplant positioner, the chuck, the imager, the cutter, and the tissuesample transporter to control and operate the plant positioner tomanipulate the individual plant for identification by the imager andposition the plant so that the sample location is proximate to thechuck, the control system further configured to operate the chuck tosecure the sample location to the chuck, operate the sampler to removethe tissue sample from the sampling location, and operate the tissuesample transporter to transport the tissue sample to the storageassembly.
 5. The automatic plant sampling apparatus of claim 4, whereinthe chuck comprises a vacuum chuck configured to supply a pressure lessthan an ambient pressure at a location proximate to a surface of thechuck, wherein when the plant positioner brings the sampling locationproximate to the chuck, the pressure draws the sample location to thechuck wherein the pressure flattens a portion of the sample location; orwherein the imager comprises an imaging system having at least onecamera.
 6. The automatic plant sampling apparatus of claim 4, whereinthe sampler comprises: (i) a laser cutting device; (ii) a device havinga plurality of knives configured to cut the tissue sample from thesample location; or (iii) a hole punching device configured to applycompressive pressure to remove the tissue sample from the samplelocation; or wherein the collection vessel comprises a microtiter havinga plurality of isolated locations each location having a scannable labelassociated with the individual plant and a dry ice receptacle forplacing dry ice proximate to the microtiter to maintain the microtiterat a pre-determined temperature.
 7. A method for automatically samplingindividual plants from a group of one or more plants, the methodcomprising: (a) loading flats containing a plurality of plants into anautomatic sampling device having a plant handling system and a plantsampling system, each plant having a unique identifier; (b) selecting anindividual plant from the tray and securing the individual plant in theplant handling system; (c) moving the individual plant with the planthandling system to the plant sampling system; (d) securing theindividual plant with the plant sampling system and releasing theindividual plant with the plant handling system; (e) identifying atleast one of a color, size, and shape of the individual plant with theplant sampling system; (f) determining a sampling location of theindividual plant based on at least one of the color, size, and shape ofthe individual plant; (g) securing the individual plant for samplingwith the plant sampling system; (h) isolating a tissue sample from theindividual plant with the plant sampling system; (i) transporting thetissue sample to a collection vessel with the plant sampling system andassociating the tissue sample with the identifier of the individualplant; and (j) storing the tissue sample for testing and returning theindividual plant for further cultivation with the plant handling system.8. The method of claim 7, wherein: step (b) comprises engaging theindividual plant with a popper of the plant handling system to move theindividual plant from a tray table of the plant handling system into agripper of the plant handling system, or wherein step (e) comprisesimaging the individual plant with an imaging system of the plantsampling system; or wherein in response to the determination of shape,size, and color of the individual plant, step (f) comprises selectingthe sampling location based on a predetermined size, color, and shapethat corresponds with desired characteristics comprising a size, color,and shape associated with an increased likelihood of plant survival; orwherein step (g) comprises orienting the individual plant so that thesample location is proximate to a chuck of the plant sampling system;positioning the sample location with the chuck so that the samplelocation is disposed for isolating; and inhibiting movement of thesample location with the chuck; or wherein step (h) comprises cuttingthe sample location with a laser cutting apparatus to remove the tissuesample from the individual plant; or wherein step (i) comprisescollecting the tissue sample with a vacuum system and carrying thetissue sample to a tissue sample collection plate associated with theindividual plant.
 9. The method of claim 8, wherein step (g) furthercomprises applying a pressure lower than an ambient pressure of theplant sampling system to draw the sample location to the chuck.
 10. Themethod of claim 7, further comprising, repeating steps (b)-(j) untileach individual plant of the plurality of plants is sampled.